Responsive membranes have been used to construct smart biomaterial interfaces. We report a novel approach to fabricate honeycomb films with a pattern of thermoresponsive polymer, namely poly(N-isopropylacrylamide). The approach was based on a combination of the breath figure method and reversible addition-fragmentation chain transfer. The hybrid film had morphological and chemical patterns resulting in varied wettability and morphology at various stages, as well as high thermo-responsiveness. Enhanced cell adhesion was observed at an incubation temperature of 37 °C, which is above its lower critical solution temperature (LCST). Furthermore, cells could be harvested at temperatures below the LCST without trypsin treatment. The non-invasive characteristics give this membrane potential as a substrate for cell sheet engineering.